Figure 4
Enhanced expression of activation markers on colonic immune cell subsets and altered colonic cytokine expression in CCR7−/− mice. (a) Colonic lamina propria (LP) leukocyte (LPL)-derived T-cell subsets (gated on CD4+ or CD8+) of 8–12-week-old CCR7−/− (n=4) and Wt mice (n=4) were stained for CD69 expression (black curve) and analyzed by flow cytometry (shaded curve: isotype control). A representative example of each group is shown. (b) Percentages of colonic LPL-derived activated CD69+ CD44high T cells of Wt (black bars) and CCR7−/− (open bars) mice (n=4 per group) are shown. Bars represent means±s.d. (c) Determination of interleukin (IL)-1β secretion from the transverse (left panels; n=10 per group) and the distal (right panels; n=5 per group) part of the colon of 8–12-week-old Wt and CCR7−/− mice. Colonic sections were cultured for 22 h in RPMI 1640 medium w/o serum at 37 °C. IL-1β release was measured in supernatants by cytokine bead array and normalized to colon weight. (d) Colon sections of Wt and CCR7−/− mice were double stained for CD11c+ cells (green) and IL-1β-expressing cells (red). Shown are representative stainings of colonic lymphoid follicular aggregates in 2 out of 3–4 mice analyzed per group. Bar=100 μm. The total numbers of colonic LPL-derived CD11c+ dendritic cells (DCs) were quantified by flow cytometry (Wt: black squares, n=3; CCR7−/−: open circles, n=3). (e) Determination of IL-15 secretion from the transverse (left panels; n=5 per group) and distal (right panels; n=5 per group) parts of the colon of 8–12-week-old Wt and CCR7−/− mice. Colonic sections were cultured for 22 h in RPMI 1640 medium w/o serum at 37 °C. IL-15 release was measured in supernatants by enzyme-linked immunosorbent assay. *P<0.05, **P<0.01, ***P<0.001; Mann–Whitney test.
